Propagation method for directly inducing bulblet from lilium longiflorum aseptic seedling leaf
A technology for small bulbs and sterile seedlings, which is applied in the field of plant tissue culture, can solve the problems of degeneration of seed characteristics, easy death of test-tube seedlings, etc., and achieves the effect of uniform size.
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Embodiment 1
[0020] Induction of lily scales: select 'Siberia' lily scales purchased from Beijing Greener Co., Ltd. without pests and diseases, wash them with detergent, rinse them under tap water for 30 minutes, soak them in 75% alcohol for 35 seconds, rinse them twice with sterile water, and then Sterilize with 0.1% mercuric chloride for different times, that is, the outer scales are sterilized for 15 minutes, the middle scales are sterilized for 12 minutes, and the inner scales are sterilized for 10 minutes, rinsed with sterile water for 4 to 5 times, and then cut in a culture dish with sterile filter paper. block spare.
[0021] Inoculate the above surface sterilized lily scales to MS+6-BA2.0mg / L+NAA0.5mg / L; MS+6-BA2.0mg / L+NAA 0.1mg / L; MS+6-BA2.0mg / L L+2,4-D 0.5mg / L; MS+6-BA2.0mg / L+2,4-D 0.1mg / L; MS+6-BA2.0mg / L+IAA0.1mg / L. Wherein, in the above medium, the sucrose content is 30g / L, and the agar content is 6.25g / L.
[0022] The results showed that the highest rate of adventitious bud ...
Embodiment 2
[0026] The clustered buds differentiated on the scales were cut into single buds and inoculated into the following eight media:
[0027] A: MS+6-BA1.0mg / L+NAA0.1mg / L;
[0028] B: MS+6-BA1.0mg / L+NAA0.5mg / L;
[0029] C: MS+6-BA1.5mg / L+NAA0.1mg / L;
[0030] D: MS+6-BA1.5mg / L+NAA0.5mg / L;
[0031] E: MS+6-BA2.0mg / L+NAA0.1mg / L;
[0032] F: MS+6-BA2.0mg / L+NAA0.5mg / L;
[0033] G: MS+6-BA2.5mg / L+NAA0.1mg / L;
[0034] H: MS+6-BA2.5mg / L+NAA0.5mg / L.
[0035] Wherein, in the above medium, the sucrose content is 30g / L, and the agar content is 6.25g / L.
[0036] The growth coefficient of clustered buds on A medium was 2.76, and the seedlings grew well; the growth coefficient of clustered shoots on B medium was 3.17, and the growth of seedlings was good; the growth coefficient of clustered shoots on C medium was 2.97, and the growth of seedlings was poor; The growth coefficient of clustered shoots on medium D was 3.96, and the seedlings were growing well; the growth coefficient of cluster...
Embodiment 3
[0039] Screening of culture medium for direct induction of bulblets from aseptic seedling leaves: Select the vigorously growing aseptic seedlings of ‘Siberia’, cut the leaves into leaf discs of about 3 cm, and inoculate them into the following four kinds of media, where the culture container is a petri dish:
[0040] MS+Pic (pyrazidine) 2.0mg / L+ sucrose 30g / L+ agar 6.25g / L;
[0041] MS+Pic (pyrazidine) 2.0mg / L+KT0.5mg / L+sucrose 30g / L+agar 6.25g / L;
[0042] MS+6-BA2.0mg / L+NAA 0.1mg / L+sucrose 30g / L+agar 6.25g / L;
[0043] MS+6-BA2.0mg / L+NAA 0.5mg / L+sucrose 30g / L+agar 6.25g / L.
[0044] Place the petri dish inoculated with leaf discs in a cardboard box, and then place the cardboard box in a completely dark artificial climate box. During the whole process, the culture temperature of the tissue culture seedlings is 20-23°C.
[0045]Among them, on the culture medium MS+Pic (pyrazidine) 2.0mg / L+sucrose 30g / L+agar 6.25g / L, the induction rate of single buds was 2.77%, while the cultur...
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